Status of large-scale analysis of post-translational modifications by mass spectrometry.
Status of large-scale analysis of post-translational modifications by mass spectrometry.
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DOI：
10.1074/mcp.o113.034181
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发表时间：
2013-12
期刊：
影响因子：
--
通讯作者：
Mann M
中科院分区：
文献类型：
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作者：
Olsen JV;Mann M
细胞功能可以通过基因表达程序来控制，但通常蛋白质翻译后修饰（PTM）提供了更精确和优雅的机制。特定修饰事件的关键功能作用-例如，在细胞周期中-已经知道了几十年，但只有在过去的10年中，基于质谱（MS）的蛋白质组学才开始揭示PTM宇宙的真实程度。在《分子和细胞蛋白质组学》PTM特刊的这篇概述中，我们评估了基于MS的蛋白质组学在大规模蛋白质修饰分析中的地位。对于许多PTM，包括磷酸化、泛素化、糖基化和乙酰化，现在可以确信地在蛋白质序列中鉴定和定位数万个位点。同样建立了不同细胞状态之间PTM水平的定量，无标记方法显示出特别的前景。它也成为可能，以确定大规模的PTM网站的绝对占有率或化学计量。已经开发了用于数千个PTM位点的生物信息学分析的强大软件。然而，尚未为任何PTM建立完整的场地清单，这种情况将持续到可预见的未来。此外，尽管基于MS的方法的PTM覆盖率令人印象深刻，但仍需要改进，特别是在组织和临床相关系统中。该领域的核心挑战是开发简化的方法来确定现在已知存在的无数修饰的生物功能。
Cellular function can be controlled through the gene expression program, but often protein post-translational modifications (PTMs) provide a more precise and elegant mechanism. Key functional roles of specific modification events—for instance, during the cell cycle—have been known for decades, but only in the past 10 years has mass-spectrometry-(MS)-based proteomics begun to reveal the true extent of the PTM universe. In this overview for the special PTM issue of Molecular and Cellular Proteomics, we take stock of where MS-based proteomics stands in the large-scale analysis of protein modifications. For many PTMs, including phosphorylation, ubiquitination, glycosylation, and acetylation, tens of thousands of sites can now be confidently identified and localized in the sequence of the protein. The quantification of PTM levels between different cellular states is likewise established, with label-free methods showing particular promise. It is also becoming possible to determine the absolute occupancy or stoichiometry of PTM sites on a large scale. Powerful software for the bioinformatic analysis of thousands of PTM sites has been developed. However, a complete inventory of sites has not been established for any PTM, and this situation will persist into the foreseeable future. Furthermore, although PTM coverage by MS-based methods is impressive, it still needs to be improved, especially in tissues and in clinically relevant systems. The central challenge for the field is to develop streamlined methods for determining biological functions for the myriad of modifications now known to exist.